Purification and characterization of vanilla bean (Vanilla planifolia Andrews [bêta]-D glucosidase

Vanilla bean [bêta]-D-glucosidase was purified to apparent homogeneity by successive anion exchange, hydrophobic interaction, and siz-exclusion chromatography. The enzyme is a tetramer (201 kDa) made up of four identical subunits (50 kDa). The optimum pH was 6.5, and the optimum temperature was 40°C at pH 7.0. Km values for [rô]-nitrophenyl-[bêta]-D-glueopyranoside and glucovanillin were 1.1 and 20.0 mM, respectively; Vmax values were 4.5 and 5.0 [mu]kat-mg-1. The [bêta]-D-glucosidase was competitively inhibited by glucono-[delta]-lactone and 1-deoxynojirimycin, with respective K. values of 670 and 152 [mu]M, and not inhibited by 2 M glucose. The [bêta]-D-glucosidase was not inhibited by [nu]-ethylmaleimide and DTNB and fully inhibited by 1.5-2 M2-mercaptoethanol and 1,4-dithiothreitol. The enzyme showed decreasing activity on [rô]-nitrophenyt-[bêta]-D-fucopyranoside, [rô]-nitrophenyl-[bêta]-Dglucopyranoside, [rô]-nitrophenyl-[bêta]-D-galactopyranoside, and [rô]-nitrophenyl-[bêta]-D-xylopyranoside. The enzyme was also active on prunasin, esculin, and salicin and inactive on cellobiose, gentiobiose, amygdalin, phloridzin, indoxyl-[bêta]-D-glucopyranoside, and quercetin-3-[bêta]-D-glucopyranoside.

Bibliographic Details

Main Authors:	Odoux, Eric, Chauwin, Audrey, Brillouet, Jean-Marc
Format:	article biblioteca
Language:	eng
Subjects:	Q02 - Traitement et conservation des produits alimentaires, Vanilla planifolia, vanille, graine, beta glucosidase, purification, vanilline, hydrolyse enzymatique, enzyme, température, concentration, activité enzymatique, http://aims.fao.org/aos/agrovoc/c_8156, http://aims.fao.org/aos/agrovoc/c_15126, http://aims.fao.org/aos/agrovoc/c_6946, http://aims.fao.org/aos/agrovoc/c_29494, http://aims.fao.org/aos/agrovoc/c_6377, http://aims.fao.org/aos/agrovoc/c_28610, http://aims.fao.org/aos/agrovoc/c_27512, http://aims.fao.org/aos/agrovoc/c_2603, http://aims.fao.org/aos/agrovoc/c_7657, http://aims.fao.org/aos/agrovoc/c_1801, http://aims.fao.org/aos/agrovoc/c_2604,
Online Access:	http://agritrop.cirad.fr/515690/ http://agritrop.cirad.fr/515690/1/515690.pdf
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